HAL Id: hal-02365027
https://hal-normandie-univ.archives-ouvertes.fr/hal-02365027
Submitted on 15 Nov 2019
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Physiological modulations of the pathogen Enterococcus faecalis MMH594 by epinephrine
Mélyssa Cambronel, Sophie Rodrigues, Florian Fécamp, Olivier Maillot, Marc Feuilloley, Nathalie Connil
To cite this version:
Mélyssa Cambronel, Sophie Rodrigues, Florian Fécamp, Olivier Maillot, Marc Feuilloley, et al.. Phys- iological modulations of the pathogen Enterococcus faecalis MMH594 by epinephrine. SFM, Oct 2018, Paris, France. �hal-02365027�
Mélyssa Cambronel, Sophie Rodrigues, Florian Fécamp, Olivier Maillot, Marc Feuilloley, Nathalie Connil
Laboratory of Microbiology Signals and Microenvironnement LMSM EA 4312, Normandie Université, Univ. Rouen Normandie, Evreux, France. www.lmsm-lab.fr
Contact: melyssa.cambronel@etu.univ-rouen.fr
Conclusion
Introduction
Nowadays, hospital-acquired infections are a huge problem of public health in the world. According to the last InVS report, several micro-organisms can induce these infections as Escherichia coli (for 23.6% of the nosocomial cases), Staphylococcus aureus (13.8%), Enterococcus faecalis (6.5%), Pseudomonas aeruginosa (6.3%), and Klebsiella pneumonia (5.6%). E. faecalis has been incriminated in endocarditis, bacteremia and urinary tract infection. It is also a bacterium which belongs to the commensal human gut microbiota, where a multitude of molecules are secreted and needed for the proper functioning of the body. Bacteria are constantly in contact with these substances, some of them can respond to these signals, by adapting their physiology and pathogenicity. This is the case of the bacteria Escherichia coli O157:H7, which is able to modulate its capacity to form biofilm and its mobilities as well as Vibrio harveyi, under exposure to epinephrine and/or norepinephrine. Bacteria are able to respond to these molecules thanks to two component systems, like QseC/QseB in E. coli O157:H7 or the BasS/BasR system in Salmonella Typhimurium. Here, we study the effect of epinephrine on growth, biofilm formation, adhesion on abiotic and biotic surfaces, hydrophobicity, autoaggregation and cytotoxicity of E. faecalis MMH 594.
Supported by :
Methods
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Physiological modulations of the pathogen
Enterococcus faecalis MMH594 by epinephrine
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References
Incubation 4h BHI, 37°C
Cultures
+/- Epinephrine (1 µM)
Adhesion and biofilm on abiotic surface
In BHI + glucose
Adhesion and cytotoxic
assay on Caco2/TC7
Karavalos, M. et al., 2012 “Pathogen espionage: multiple bacterial adrenergic sensors eavesdrop on host communication systems” Molecular Microbiology, vol.
87, no. 3, p455-465
Lesouhaitier O. et al., 2009 “Gram-negative bacterial sensors for eukaryotic signal molecules” Sensors, vol. 9, no. 9, p6967-6990
Bansal T. et al., 2007 “Differential effects of epinephrine , norepinephrine, and indole on Escherichia coli O157:H7 chemotaxis, colonization, and gene expression”
Infection and Immunity, vol. 75, no. 9, p4597-4607
Yang. Q et al., 2014 “Norepinephrine and dopamine increase motility, biofilm formation, and virulence of Vibrio harveyi” Frontiers in Microbiology, vol. 5
Enquête de prévalence des infections nosocomiales et des traitements anti-infectieux en établissement de santé, France, mai-juin 2012 , Institut de veille sanitaire Enquête de prévalence des infections nosocomiales et des traitements anti-infectieux en établissement de santé, France, mai-juin 2017 , Institut de veille sanitaire
Hydrophobicity and autoagreggation assay Overnight Cultures of MMH 594
Bacterial growth assay
NS **
* NS
0%
20%
40%
60%
80%
100%
120%
140%
160%
Adhesion (% of control)
Control 100 µM Epi 10 µM Epi 1 µM Epi 0.1 µM Epi NS
NS
NS
NS
0%
20%
40%
60%
80%
100%
120%
140%
160%
Biofilm (% of control)
Control 100 µM Epi 10 µM Epi 1 µM Epi 0.1 µM Epi
Biofilm and adhesion on abiotic and biotic surfaces Hydrophobicity and autoaggregation assay
Cytotoxic activity/viability assay on Caco2/TC7 cells
-> The pre-treatment of MMH 594 with 1 µM of epinephrine leads to an increase of its cytotoxicity (reduction of the viable Caco2/TC7 cells)
-> No impact of 1 µM of epinephrine on these parameters.
Parameters related to the capacity of adhesion of E. faecalis.
-> The capacity of MMH 594 to adhere on abiotic surface is increased upon exposure to 10 µM and 1 µM of epinephrine.
Epinephrine does not modify its capacity to form biofilm or its adhesion towards the intestinal Caco2/TC7 cell line.
Epinephrine 100 , 10, 1 and 0,1 µM
0%
20%
40%
60%
80%
100%
120%
Viable cells (% of control)
Control Triton MMH 594 MMH 594 / 1µM Epi
Putative two component systems (TCS) homologous to epinephrine’s sensor
-> Exposure to 1 µM of epinephrine does not modify the growth curve of the strain
E. faecalis MMH 594.
This study showed that epinephrine can impact some aspects of Enterococcus faecalis MMH 594. One micromolar of epinephrine does not modify its growth curve but can modulate its capacity to adhere on abiotic surface and its cytotoxicity towards the intestinal cell line Caco2/TC7. Epinephrine can also, at a concentration of 10 µM, increase its adhesion to abiotic surface. However, the biofilm is not impacted upon exposure to various concentrations of epinephrine as well as hydrophobicity and autoagreggation. Complementary studies like biofilm including an adhesion step are necessary, along with qRT-PCR of genes involved in adhesion.
The strain MMH 594 contains many putative two component systems (TCS) that may be interesting candidates to identify the sensor able to perceive epinephrine. Molecular docking and mutants construction could help to find the sensor of epinephrine in this particular strain.
0,01 0,1 1 10
0 5 10 15 20 25 30
LOG (OD 580)
Time (hours) Control 1 µM Epi
NS
0%
20%
40%
60%
80%
100%
120%
140%
160%
Adhesion on Caco2/TC7 (% of control)
Control 1 µM Epi
NS
0,00 10,00 20,00 30,00 40,00 50,00 60,00 70,00
% of hydrophobicity
Control 1 µM Epi
NS
0,00 10,00 20,00 30,00 40,00 50,00 60,00 70,00 80,00 90,00 100,00
% of autoaggregation
Control 1 µM Epi
*
EOI84084.1 EOI84085.1
S. Typhimurium
EOI80856.1
33% 30%
38% 27%
EOI85394.1 EOI85393.1
S. Typhi
EOI81342.1 EOI81341.1
33% 36%
EOI80857.1 EOI80856.1
27% 36%
30% 40%
EOI80857.1
-> The two component system EOI80856.1/EOI80857.1 is homologous to the four two component systems known to sense epinephrine in three different bacteria.
Each sequence of TCS known to play a role in the recognition of epinephrine by three specific bacteria were blasted against the genome of the strain MMH 594. For each system known, one or more putative TCS from
MMH 594 are
represented.
basR basS
qseB qseC
E. coli O157:H7
EOI80857.1 EOI80856.1
29% 33%
MMH 594
E. coli O157:H7
EOI80856.1
27% 36%
EOI80857.1
qseF qseE
MMH 594
MMH 594
MMH 594
Response regulator (RR) Sensor
Homologous RR Homologous sensor
cpxR cpxA
